The Development of Biosensor with Imaging Ellipsometry

The concept of biosensor with imaging ellipsometry was proposed about ten years ago. It has become an automatic analysis technique for protein detection with merits of label-free, multi-protein analysis, and real-time analysis for protein interaction process, etc. Its principle, andrelated technique units, such as micro-array, micro-fluidic and bio-molecule interaction cell, sampling unit and calibration for quantitative detection as well as its applications in biomedicine field are presented here.

An auto-focusing method for imaging ellipsometry system

An auto-focusing method based on the image brightness gradient sharpness function is presented for imaging ellipsometry system, in which the image plane of the thin-film specimen is not perpendicular to the optical axis. The clear image of a specimen with large area is obtained by moving the imaging sensor in optical axis direction and around its sensitive surface centre successively. The experimental results demonstrate its feasibility.

An automatic imaging spectroscopic ellipsometer for characterization of nano-film pattern on solid substrate

In order to characterize the physical and spatial properties of nano-film pattern on solid substrates, an automatic imaging spectroscopic ellipsometer (ISE) based on a polarizer - compensator - specimen - analyzer configuration in the visible region is presented. It can provide the spectroscopic ellipsometric parameters psi (x, y, lambda) and Delta (x, y, lambda) of a large area specimen with a lateral resolution in the order of some microns. A SiO2 stepped layers pattern is used to demonstrate the function of the ISE which shows potential application in thin film devices' such as high-throughput bio-chips.

Phage M13Ko7 Detection With Biosensor Based On Imaging Ellipsometry And Afm Microscopic Confirmation

A rapid detection and identification of pathogens is important for minimizing transfer and spread of disease. A label-free and multiplex biosensor based on imaging ellipsometry (BIE) had been developed for the detection of phage M13KO7. The surface of silicon wafer is modified with aldehyde, and proteins can be patterned homogeneously and simultaneously on the surface of silicon wafer in an array format by a microfluidic system. Avidin is immobilized on the surface for biotin-anti-M13 immobilization by means of interaction between avidin and biotin, which will serve as ligand against phage M13KO7. Phages M13KO7 are specifically captured by the ligand when phage M13KO7 solution passes over the surface, resulting in a significant increase of mass surface concentration of the anti-M13 binding phage M13KO7 layer, which could be detected by imaging ellipsometry with a sensitivity of 10(9) pfu/ml. Moreover, atomic force microscopy is also used to confirm the fact that phage M13KO7 has been directly captured by ligands on the surface. It indicates that BIE is competent for direct detection of phage M13KO7 and has potential in the field of virus detection. (C) 2008 Elsevier B.V. All rights reserved.

Detection of hepatitis B virus markers using a biosensor based on imaging ellipsometry

A biosensor based on imaging ellipsometry (BIE) has been developed and validated in 169 patients for detecting five markers of hepatitis B virus (HBV) infection. The methodology has been established to pave the way for clinical diagnosis, including ligand screening, determination of the sensitivity, set-up of cut-off values (CoVs) and comparison with other clinical methods. A matrix assay method was established for ligand screening. The CoVs of HBV markers were derived with the help of receiver operating characteristic curves. Enzyme-linked immunosorbent assay (ELISA) was the reference method. Ligands with high bioactivity were selected and sensitivities of 1 ng/mL and 1 IU/mL for hepatitis B surface antigen (HBsAg) and surface antibody (anti-HBs) were obtained respectively. The CoVs of HBsAg, anti-HBs, hepatitis B e antigen, hepatitis B e antibody and core antibody were as follows: 15%, 18%, 15%, 20% and 15%, respectively, which were the percentages over the values of corresponding ligand controls. BIE can simultaneously detect up to five markers within 1 h with results in acceptable agreement with ELISA, and thus shows a potential for diagnosing hepatitis B with high throughput.

Imaging properties of coherent anti-Stokes Raman scattering microscope

The coherent anti-Stokes Raman scattering (CARS) microscope with the combination of confocal and CARS techniques is a remarkable alternative for imaging chemical or biological specimens that neither fluoresce nor tolerate labelling. CARS is a nonlinear optical process, the imaging properties of CARS microscopy will be very different from the conventional confocal microscope. In this paper, the intensity distribution and the polarization property of the optical field near the focus was calculated. By using the Green function, the precise analytic solution to the wave equation of a Hertzian dipole source was obtained. We found that the intensity distributions vary considerably with the different experimental configurations and the different specimen shapes. So the conventional description of microscope (e.g. the point spread function) will fail to describe the imaging properties of the CARS microscope.

Heterodimerization of integrin Mac-1 subunits studied by single-molecule imaging

Heterodimerization of integrin Mac-1 (alpha(M) beta(2)) Subunits plays important role on regulating leukocytes adhesion to extracellular matrix or endothelial cells. Here, using total internal reflection microscopy, we investigated the heterodimerization of integrin Mac-1 subunits at the single-molecule level in live cells. Individual alpha(M) subunit fused to the enhanced yellow fluorescent protein (eYFP) was imaged at the basal plasma membrane of live Chinese hamster ovary (CHO) cells. Through analysis of mean square displacement (MSD), diffusion coefficient, the size of restricted domain and fraction of molecules undergoing restricted diffusion, we found that as compared with the diffusion in the absence of beta(2) subunit, the diffusion of single-molecule of alpha(M)-YFP was suppressed significantly in the presence of beta(2) subunit. Thus, based on the oligomerization-induced trapping model, we suggested that in the presence of beta(2) subunit, the am subunit may form heterodimer with it. (C) 2008 Elsevier Inc. All rights reserved.

激光光束实时监测与自动准直系统设计

设计了一个激光光斑实时监测与光路自动准直装置,能够实时监测激光光斑并自动准直激光输出方向。基于透镜成像原理,使用CCD探测器获得光斑的二维成像,并根据两点确定一条直线原理和使用压电陶瓷电动调整架实现光路自动准直;监测控制程序采用虚拟仪器开发软件Lab View编写,可以实时监测激光光斑模式与光斑位置抖动情况,并进行反馈控制。经测试,设计装置的调整精度达0.5μrad,反馈控制频率约1 Hz,完全可降低或消除抖动周期在1 s以上的光斑飘移。

Near-field x-ray phase contrast imaging and phase retrieval algorithm

Theoretical analyses of x-ray diffraction phase contrast imaging and near field phase retrieval method are presented. A new variant of the near field intensity distribution is derived with the optimal phase imaging distance and spatial frequency of object taken into account. Numerical examples of phase retrieval using simulated data are also given. On the above basis, the influence of detecting distance and polychroism of radiation on the phase contrast image and the retrieved phase distribution are discussed. The present results should be useful in the practical application of in-line phase contrast imaging.

Phase imaging with partially coherent x rays

X-ray phase imaging with illumination by a partially coherent source with a setup similar to in-line holography is considered. Using the optical transform function, we consider the effects of partial coherence on this x-ray phase imaging for a weak phase object. The optimal contrast and the resolution of phase imaging are analyzed. As the coherence decreases, the imaging contrast and the optimal contrast frequency decrease, and the resolution degrades. It is shown that this contrast-enhanced phase-imaging method can be regarded as a linear bandpass filter and that the bandwidth and the image contrast are changeable. The frequency property of the imaging system can be improved if an incoherent x-ray source with the proper shape is used. (C) 1999 Optical Society of America.

Instability of solution of phase retrieval in direct diffraction phase-contrast imaging with partially coherent X-ray source

The theoretical model of direct diffraction phase-contrast imaging with partially coherent x-ray source is expressed by an operator of multiple integral. It is presented that the integral operator is linear. The problem of its phase retrieval is described by solving an operator equation of multiple integral. It is demonstrated that the solution of the phase retrieval is unstable. The numerical simulation is performed and the result validates that the solution of the phase retrieval is unstable.

Resolution and noise in ghost imaging with classical thermal light

The resolution and classical noise in ghost imaging with a classical thermal light are investigated theoretically. For ghost imaging with a Gaussian Schell model source, the dependences of the resolution and noise on the spatial coherence of the source and the aperture in the imaging system are discussed and demonstrated by using numerical simulations. The results show that an incoherent source and a large aperture will lead to a good image quality and small noise.

Cross spectral purity and its influence on ghost imaging experiments

Pseudo-thermal light has been widely used in ghost imaging experiments. In order to understand the differences between the pseudo-thermal source and thermal source, we propose a method to investigate whether a light source has cross spectral purity (CSP), and experimentally measure the cross spectral properties of the pseudo-thermal light source in near-field and far-field zones. Moreover we present a theoretical analysis of the cross spectral influence on ghost imaging. (c) 2006 Elsevier B.V. All rights reserved.

Transmission area and correlated imaging

The relationship between transmission area of an object imaged and the visibility of correlated imaging is investigated in a lensless system. We show that they are not in simple inverse proportion, as usually depicted. The changes of the visibility will be quite different when the transmission area is varied by different manners, which may motivate people to seek a new understanding about the influence factors of the visibility. (C) 2007 Optical Society of America

Electromagnetically induced self-imaging

We study the self-imaging and image-transforming properties of a probe field in a cold atomic medium with electromagnetically induced transparency (EIT). Due to the similarities between the gradient-index medium and the inhomogeneous index distribution of an EIT medium under the conditions of a negative probe detuning and a Gaussian control field, we find based on analytical investigations that there exists a kind of electromagnetically induced self-imaging phenomenon in cold atomic media. Numerical simulations clearly show that electromagnetically induced self-imaging is observable and controllable. (c) 2007 Optical Society of America.